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Replication data for: A novel sweet potato potyvirus open reading frame (ORF) is expressed via polymerase slippage and suppresses RNA silencing

International Potato Center Dataverse OAI Archive

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Title Replication data for: A novel sweet potato potyvirus open reading frame (ORF) is expressed via polymerase slippage and suppresses RNA silencing
 
Identifier https://doi.org/10.21223/P3/3AZYQO
 
Creator Untiveros, Milton
Olspert, Allan
Artola, Katrin
Firth, Andrew
Kreuze, Jan
Valkonen, Jari
 
Publisher International Potato Center
 
Description The accumulation of GFP mRNA and GFP mRNA-derived siRNA was evaluated by northern blot analysis in leaf tissues agroinfiltrated for coexpression of GFP and P1, P1N-PISPO, SPFMV HCpro, SPLV P1, PVA HCpro or GUS. Total RNA was extracted with TRIzol LS Reagent (Invitrogen) according to the manufacturer’s recommendations alow-molecular-weight (LMW) and high-molecular-weight (HMW) RNA fractions were separated as described in Cuellar et al. (2009). LMW RNA (15 lg/lane) and HMW RNA (10 lg/lane) were separated by electrophoresis in a denaturing 15% polyacrylamide gel and 1.2% (w/v) agarose gel, respectively, both containing formaldehyde. The two gels were transferred to a nylon membrane (GE Healthcare) and crosslinked chemically with 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide (Pall and Hamilton, 2008) or by UV light (Sambrook and Russell, 2001), respectively. A GFP mRNA-specific radioactive RNA probe was synthesized, fractionated with carbonate hydrolysis and used to probe the membranes for GFP mRNA and GFP mRNA-derived siRNA, as described previously (Cuellar et al., 2009). Signals were detected by autoradiography on an X-ray film (Kodak)
 
Subject Agricultural Sciences
Frameshift
Gene expression
Overlapping gene
RNA silencing suppression
RNA virus
Transcriptional slippage
 
Language English
 
Contributor Administrator, CIP
CGIAR Research Program on Roots, Tubers and Bananas (RTB)
International Potato Center
Fuentes, Segundo
 
Type fastq